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Usefulness of real time PCR to quantify parasite load in serum samples from chronic Chagas disease patients

Overview of attention for article published in Parasites & Vectors, March 2015
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87 Mendeley
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Title
Usefulness of real time PCR to quantify parasite load in serum samples from chronic Chagas disease patients
Published in
Parasites & Vectors, March 2015
DOI 10.1186/s13071-015-0770-0
Pubmed ID
Authors

Myllena F Melo, Otacilio C Moreira, Priscila Tenório, Virginia Lorena, Izaura Lorena-Rezende, Wilson Oliveira Júnior, Yara Gomes, Constança Britto

Abstract

Inconclusive results of serological diagnosis in Chagas disease have an important impact on blood banks worldwide, reflecting in the high number of discarded bags or in an increased transmission by blood transfusion. Molecular techniques such as qPCR have been used for diagnosis and to monitor Trypanosoma cruzi load in peripheral blood samples. A promising perspective refers to the possibility of parasite DNA detection in serum, taking advantage in using the same samples collected for serological screening. In order to evaluate the effectiveness of a qPCR strategy for detecting and quantifying T. cruzi DNA in serum, we selected 40 chronic Chagas disease patients presenting different clinical manifestations: Cardiac (23), Digestive (4), Mixed form [cardiodigestive] (7), and asymptomatic (6). Twenty seronegative individuals from non-endemic areas were included as controls. Samples were extracted using QIAamp DNA mini kit (QIAGEN) and qPCR was performed in a multiplex format with TaqMan probes for the nuclear satellite DNA of T. cruzi and for the human RNase P gene. In addition, DNA migration to serum during blood coagulation was assessed using a commercial exogenous control (Exo IPC, Applied Biosystems) in a separate qPCR reaction. The comparative duplex qPCR analysis revealed that, even with an increase in Ct values, it was possible to detect all DNA targets in serum. In addition, the same linearity range for T. cruzi quantification (from 10(5) to 0.5 par. eq./mL) between serum, blood or culture samples (T. cruzi epimastigotes - Cl Brener strain) was found. When patient samples were evaluated, no significant differences in parasite load between the distinct clinical manifestations were found for both blood and serum samples. Moreover, median values of parasite burden were 1.125 and 1.230 par. eq./mL for serum and blood, respectively. Using serology as gold standard, we found 95% sensitivity for T. cruzi detection in serum and 97.5% for blood, and 100% specificity for both samples. Taken together, our data indicate the potential of using serum samples for molecular diagnosis and parasite load quantification by qPCR, suggesting its use in reference laboratories for the diagnosis of Chagas disease patients.

Twitter Demographics

The data shown below were collected from the profiles of 2 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 87 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 87 100%

Demographic breakdown

Readers by professional status Count As %
Student > Master 17 20%
Researcher 14 16%
Student > Ph. D. Student 12 14%
Student > Doctoral Student 10 11%
Other 5 6%
Other 12 14%
Unknown 17 20%
Readers by discipline Count As %
Agricultural and Biological Sciences 20 23%
Biochemistry, Genetics and Molecular Biology 14 16%
Medicine and Dentistry 10 11%
Immunology and Microbiology 4 5%
Engineering 3 3%
Other 12 14%
Unknown 24 28%

Attention Score in Context

This research output has an Altmetric Attention Score of 2. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 19 November 2015.
All research outputs
#12,067,540
of 19,156,856 outputs
Outputs from Parasites & Vectors
#2,624
of 4,891 outputs
Outputs of similar age
#200,439
of 386,060 outputs
Outputs of similar age from Parasites & Vectors
#245
of 500 outputs
Altmetric has tracked 19,156,856 research outputs across all sources so far. This one is in the 36th percentile – i.e., 36% of other outputs scored the same or lower than it.
So far Altmetric has tracked 4,891 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 5.0. This one is in the 45th percentile – i.e., 45% of its peers scored the same or lower than it.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 386,060 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 47th percentile – i.e., 47% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 500 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 50% of its contemporaries.